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US 12,492,380 B2Granted
Methods for the production of hepatocytes
AssigneesFUJIFILM Cellular Dynamics, Inc., FUJIFILM Holdings America Corporation
InventorsIgor Gurevich, Sarah Burton, Christie Munn, Madelyn Donegan, Katherine Czysz, Deepika Rajesh, Makiko Ohshima
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Abstract
The present disclosure provides methods of producing hepatocytes from induced pluripotent stem cells. Further provided herein are methods of using the hepatocytes for the treatment of a liver disease.
Claims25
A method for producing hepatocytes comprising:
- (a) culturing induced pluripotent stem cells (iPSCs) in the presence of a GSK3 inhibitor to provide pre-conditioned iPSCs;
- (b) differentiating the pre-conditioned iPSCs to definitive endoderm (DE) cells, wherein differentiating to DE cells comprises sequentially culturing the iPSCs in a first endoderm induction media (EIM) comprising Activin A, a second EIM comprising BMP4, VEGF, and bFGF, and a third EIM comprising VEGF and DMSO;
- (c) culturing the DE cells to induce formation of hepatoblasts;
- and
- (d) differentiating the hepatoblasts to hepatocytes by culturing the hepatoblasts in hepatocyte differentiation media (HDM) comprising a GSK3 inhibitor to produce the hepatocytes.
2 more elements
A composition comprising hepatocyte cells, at least 90% positive for AAT and/or at least 80% positive for albumin.
Dependencies are inferred from the claim text.